RedoxiFluor: A microplate technique to quantify target-specific protein thiol redox state in relative percentage and molar terms.

ABSTRACT: Unravelling how reactive oxygen species regulate fundamental biological processes is hampered by the lack of an accessible microplate technique to quantify target-specific protein thiol redox state in percentages and moles. To meet this unmet need, we present RedoxiFluor. RedoxiFluor uses two spectrally distinct thiol-reactive fluorescent conjugated reporters, a capture antibody, detector antibody and a standard curve to quantify target-specific protein thiol redox state in relative percentage and molar terms. RedoxiFluor can operate in global mode to assess the redox state of the bulk thiol proteome and can simultaneously assess the redox state of multiple targets in array mode. Extensive proof-of-principle experiments robustly validate the assay principle and the value of each RedoxiFluo

SUBMITTER: Tuncay A 

PROVIDER: S-EPMC8904371 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature